CAJ | 학술논문

目的探讨凝血因子Ⅶ(FⅦ)与小鼠进展性脑挫裂伤出血的关系,为重组人FⅦa的临床应用提供实验依据. 方法 (1)12只BALB/c雄性小鼠分别给予1,3,5,10 mg/kg的脂质体包裹的FⅦsiRNA注射动物尾静脉,每剂量组3只,另3只注射等渗盐水,注射后2d予眼球取血,PCR检测肝脏FⅦ的表达,ELISA法检测血浆FⅦ浓度,底物显色法检测血浆FⅦ活性;选择最佳剂量的脂质体包裹的FⅦsiRNA来抑制小鼠FⅦ的表达.(2)30只BALB/c雄性小鼠按随机数字表法分为FⅦ抑制组和对照组,FⅦ抑制组给予最佳剂量的脂质体包裹的FⅦsiRNA;对照组则给予同等剂量的脂质体包裹的阴性对照siRNA,每组15只,均建立脑挫裂伤出血模型,于伤后3,24,72 h检测两组脑挫裂伤出血量;伤后24,48 h观察两组血肿体积. 结果 (1)1,3,5,10 mg/kg脂质体包裹的FⅦsiRNA抑制后,肝脏FⅦ的表达均明显下降,血浆FⅦ浓度和活性也明显下降.FⅦsiRNA的最佳剂量为3 mg/kg.(2)FⅦ抑制组伤后3,24,72 h相对脑出血量分别为1.46±0.10,1.82 ±0.23,2.28±0.15,均显著大于对照组(1.00 ±0.25,1.20 ±0.31,1.20±0.22)(P＜0.05).伤后24,48 hFⅦ抑制组血肿体积分别为(6.7±1.5)mm3、(9.8±1.0)mm3,均较对照组[(5.2±1.2)mm3、(5.5±1.5)mm3]明显增加(P＜0.01). 结论体内FⅦ浓度与进展性脑挫裂伤出血的发生密切相关,脑挫裂伤出血后给予FⅦ可以显著降低进展性脑挫裂伤出血的发生.
목적 탐토응혈인자Ⅶ(FⅦ)여소서진전성뇌좌렬상출혈적관계,위중조인FⅦa적림상응용제공실험의거. 방법 (1)12지BALB/c웅성소서분별급여1,3,5,10 mg/kg적지질체포과적FⅦsiRNA주사동물미정맥,매제량조3지,령3지주사등삼염수,주사후2d여안구취혈,PCR검측간장FⅦ적표체,ELISA법검측혈장FⅦ농도,저물현색법검측혈장FⅦ활성;선택최가제량적지질체포과적FⅦsiRNA래억제소서FⅦ적표체.(2)30지BALB/c웅성소서안수궤수자표법분위FⅦ억제조화대조조,FⅦ억제조급여최가제량적지질체포과적FⅦsiRNA;대조조칙급여동등제량적지질체포과적음성대조siRNA,매조15지,균건립뇌좌렬상출혈모형,우상후3,24,72 h검측량조뇌좌렬상출혈량;상후24,48 h관찰량조혈종체적. 결과 (1)1,3,5,10 mg/kg지질체포과적FⅦsiRNA억제후,간장FⅦ적표체균명현하강,혈장FⅦ농도화활성야명현하강.FⅦsiRNA적최가제량위3 mg/kg.(2)FⅦ억제조상후3,24,72 h상대뇌출혈량분별위1.46±0.10,1.82 ±0.23,2.28±0.15,균현저대우대조조(1.00 ±0.25,1.20 ±0.31,1.20±0.22)(P＜0.05).상후24,48 hFⅦ억제조혈종체적분별위(6.7±1.5)mm3、(9.8±1.0)mm3,균교대조조[(5.2±1.2)mm3、(5.5±1.5)mm3]명현증가(P＜0.01). 결론 체내FⅦ농도여진전성뇌좌렬상출혈적발생밀절상관,뇌좌렬상출혈후급여FⅦ가이현저강저진전성뇌좌렬상출혈적발생.
Objective To study the correlation between the coagulation factor Ⅶ (F Ⅶ) and progressive hemorrhage after brain contusion in mice and provide the experimental evidence for the clinical application of recombinant human FⅦa.Methods Twelve male BALB/c mice were given liposomeencapsulated FⅦsiRNA via tail vein at doses of 1,3,5 and 10 mg/kg with 3 mice per dosage.The other 3 mice received equivalent volume of normal saline as controls.Two days after the injection,mice blood sampling was used to detect FⅦ mRNA expression in liver using real-time PCR,level of plasma FⅦ using ELISA method,and activity of plasma FⅦ using chromogenic substrate assay.The optimal dose at which F Ⅶ expression was inhibited was determined.Thirty BALB/c male mice were assigned to two groups (n =15 per group) according to the random number table:FⅦ-suppressing group,mice were injected with FⅦsiRNA at the optimal dose and control group,mice were injected with same volume of negative control vector.The model of brain contusion was established in both groups.Volume of hemorrhage following brain contusion was measured at 3,24 and 72 h postinjury,and hematoma volume at 24 and 48 h postinjury.Results Liposome-encapsulated siRNA delivery down-regulated FⅦ expression in the mouse liver.Level and activity of plasma FⅦ were also reduced significantly.The optimal siRNA dose was 3 mg/kg.At 3,24 and 72 h postinjury,relative volume of brain hemorrhage in FⅦ-suppressing group was 1.46 ± 0.10,1.82 ± 0.23 and 2.28 ± 0.15 respectively,significantly higher than that in control group (1.00 ± 0.25,1.20 ± 0.31 and 1.20 ± 0.22 respectively) (P ＜ 0.05).At 24 and 48 h postinju-ry,volume of hematoma in FⅦ-suppressing group was (6.7 ± 1.5)mm3 and (9.8 ± 1.0) mm3,significantly higher than that in control group [(5.2 ± 1.2) mm3 and (5.5 ± 1.5) mm3] (P ＜0.01).Conclusions Level of FⅦ in vivo relates closely to the progressive hemorrhage of brain contusion in mice.Administration of FⅦ is effective to reduce the incidence of progressive hemorrhage.